Monique Médina

The distribution of GABA-immunoreactive neurons in the brain of the silver eel (Anguilla anguilla L.).

The distribution of GABA-immunoreactivity was studied in the brain of the silver eel (Anguilla anguilla) by means of antibodies directed against GABA. Immunoreactive neuronal somata were distributed throughout the brain. Positive perikarya were detected in the internal cellular layer of the olfactory bulb, and in all divisions of the telencephalon, the highest density being observed along the midline. Numerous GABA-reactive cell bodies were found in the diencephalon, particularly in the preoptic and tuberal regions of the hypothalamus, and the dorsolateral, dorsomedial and ventromedial thalamic nuclei. In the optic tectum, the majority of GABApositive cell bodies were located in the periventricular layer. A number of immunolabelled cell bodies were observed in different tegmental structures, notably the torus semicircularis. In the cerebellum, the Purkinje cells were either very intensely or very weakly immunoreactive. In the rhombencephalon, reactive cell bodies were observed in the eminentia granularis, the valvula cerebellaris, the octavolateral nucleus, the lobus vagus and in the vagal and glossopharyngeal motor nuclei. Intensely immunoreactive axons and terminals were observed in the external granular layer and internal cellular layer of the olfactory bulb. In the telencephalon, the highest density of reactive fibres and boutons was found in the fields of the medial wall. Many immunolabelled fibres were seen in the medial and lateral forebrain bundles. In the diencephalon, intense labelling of fibres and terminals were observed in the nuclei situated close to the midline. In the optic tectum the highest density of reactive fibres was seen in the sfgs, the layer to which the retina projects massively. Finally, in the rhombencephalon the strongest labelling of neurites was observed in the nuclei of the raphé, the nucleus octavocellularis magnocellularis and the nuclei of the IXth and Xth cranial nerves. The GABAergic system of the eel, which is well developed, appears to be generally comparable to that described in tetrapod vertebrates.

GABA immunoreactivity in the nucleus isthmo-opticus of the centrifugal visual system in the pigeon: a light and electron microscopic study.

The present study examined GABA immunoreactivity within the retinopetal nucleus isthmo-opticus (NIO) of the pigeon centrifugal visual system (CVS) using light- (immunohistofluorescence, peroxidase anti-peroxidase: PAP) and electron- (postembedding GABA immunogold) microscopic techniques. In some double-labeling experiments, the retrograde transport of the fluorescent dye rhodamine beta-isothiocyanate (RITC) after its intraocular injection was combined with GABA immunohistofluorescence. GABA-immunoreactive (-ir) somata were demonstrated within the neuropilar zone of the NIO adjacent to the centrifugal cell laminae whereas the centrifugal neurons were always immunonegative. A quantitative ultrastructural analysis was performed which distinguished five categories of axon terminal profiles (P1-5) on the basis of various cytological criteria: type of synaptic contact (symmetrical or asymmetrical); shape, size, and density of synaptic vesicles as well as the immunolabeling (positive or negative), size of profile and appearance of hyaloplasm. Numerous GABA-ir afferents to centrifugal neurons via axon terminal types P2a, P2c, and P3 were observed which comprised 47.1% of the total input. Moreover, the data suggest that some of the P2a terminals, which make up 26.4% of the input, stem from the intrinsic GABA-ir interneurons, whereas the latter receive P1, P3, but also P2 terminal input, indicating that interneurons may contact other interneurons via type P2a axon terminals. The results also suggest that the GABA-ir P3 or the immunonegative P1b and P5 axon terminals are of extrinsic origin arising from cells in the optic tectum whereas the P2c and P4 axon terminals are associated with extra-tectal input to the NIO. The GABAergic innervation of centrifugal neurons within the NIO may be the basis for the demonstrated facilitatory effect of the centrifugal output upon ganglion cell responses. This is relevant to hypotheses regarding CVS involvement in attentional mechanisms through selective enhancement of retinal sensitivity depending on the location of meaningful or novel stimuli.

Evidence of GABA-immunopositive neurons in the dorsal part of the lateral geniculate nucleus of reptiles: Morphological correlates with interneurons

The distribution and staining pattern of gamma-aminobutyric acid immunoreactivity have been examined by both light and electron microscopy in the dorsal part of the lateral geniculate nucleus of three reptilian species: the turtle Chinemys reevesi, the lizard Ophisaurus apodus and the snake Vipera aspis. After perfusion of the animals with 1% paraformaldehyde and 1% glutaraldehyde and polyethyleneglycol embedding of the brains, the analysis of sections processed immunocytochemically with an anti-GABA antiserum has revealed a moderate-to-dense labeling of the neurons of the dorsal part of the lateral geniculate complex in these species. Labeled cell bodies are small-sized, either rounded or fusiform and the GABA-positive dendrites emerging from them are not preferentially oriented in any particular direction. Quantitative studies in Vipera indicate that GABA-positive neurons make up about 14% of the population of neurons of the dorsal part of the lateral geniculate nucleus. Electron microscopy of specimens treated by either pre- or post-embedding techniques has confirmed that these cells corresponded to neurons. No glial cells were ever observed to be immunopositive. These GABA-positive neurons, characterized by the presence of pleiomorphic synaptic vesicles localized either in their perikaryon or more often in presynaptic dendrites, established symmetrical synaptic contacts. In this case, the latter were involved both pre- and postsynaptically in serial and, more rarely, in triadic arrangements, a synaptic organization specific to interneurons. The involvement of such GABA-positive neurons in local circuits is discussed.

Function and Cytochemical Characteristics of Postural Limb Muscles of the Rhesus Monkey: A Telemetered EMG and Immunofluorescence Study

Using telemetered electromyography and immunocytochemical fibre typing (of both fresh frozen and preserved specimens), the present paper demonstrates clearly that at the elbow, knee, and ankle joints, the rhesus monkey (Macaca mulatta) is endowed with one extensor-muscle head specialized for posture. These postural heads are distinguished by (a) recruitment at low levels to maintain joint position against the effect of gravity, (b) recruitment near maximum levels during walking, and (c) high content and relatively large size of slow, fatigue-resistant (type I) muscle fibres. The nonpostural heads of the investigated muscles were recruited at levels correlated to the strenuousness of the effort and are notable by the small percentage and size of slow muscle fibres. The postmortem stability of the structural properties of myosin makes immunocytochemical fibre typing suitable for the study of preserved cadavers.

Retinal projections in two crocodilian species, Caiman crocodilus and Crocodylus niloticus

 The retinal projections of Caiman crocodilus and Crocodylus niloticus were investigated by means of the orthograde axonal transport of either rhodamine β-isothiocyanate or tritiated proline. In these two species, each tracer revealed contralateral retinal projections to three hypothalamic regions (subventricular gray matter, nucleus suprachiasmaticus, and area optica hypothalami lateralis), five thalamic regions (nuclei ovalis, dorsolateralis anterior, ventrolateralis and ventrobasalis, and lateral geniculate complex, of which six subcomponents can be distinguished), six pretectal regions (nuclei posterodorsalis, lentiformis mesencephali, griseus tectalis, geniculatus pretectalis, area optica commissurae posterior and area optica pretectalis lateroventralis), six outermost layers of the optic tectum, and the nucleus opticus tegmenti. Weak ipsilateral retinal projections have been observed in two hypothalamic nuclei and in the nucleus opticus tegmenti. Comparative analysis with other data show that the contralateral retinal projections of crocodiles are considerably more reptilian than avian. Moreover, crocodiles share with birds an extremely poor contingent of ipsilateral retinal projections.

Serotonergic retinopetal projections from the dorsal raphe nucleus in the mouse demonstrated by combined [3H] 5-HT retrograde tracing and immunolabeling of endogenous 5-HT

The present study demonstrated a direct serotonergic retinopetal projection in the mouse stemming from the lateral portion of the dorsal raphe nucleus bilaterally. A double-labeling technique was employed combining: (1) radioautography and retrograde axonal tracing following intraocular injection of [(3)H] 5-HT and (2) immunocytochemical identification of endogenous 5-HT. Radiolabeled neurons were only observed within the dorsal raphe nucleus and were always double-labeled with the 5-HT antibody. The radiolabeling appeared to be specific resulting from the retrograde transport of a radioactive 5-HT derivative product following uptake of the neurotransmitter by intraretinal terminals.

Centrifugal visual system of Crocodylus niloticus: A hodological, histochemical, and immunocytochemical study

The retinopetal neurons of Crocodylus niloticus were visualized by retrograde transport of rhodamine beta‐isothiocyanate or Fast Blue administered by intraocular injection. Approximately 6,000 in number, these neurons are distributed in seven regions extending from the mesencephalic tegmentum to the rostral rhombencephalon, approximately 70% being located contralaterally to the injected eye. None of the centrifugal neurons projects to both retinae. The retinopetal neurons are located in rostrocaudal sequence in seven regions: the formatio reticularis lateralis mesencephali, the substantia nigra, the griseum centralis tectalis, the nucleus subcoeruleus dorsalis, the nucleus isthmi parvocellularis, the locus coeruleus, and the commissura nervi trochlearis. The greatest number of cells (approximately 93%) is found in the nucleus subcoeruleus dorsalis. The majority are multipolar or bipolar in shape and resemble the ectopic centrifugal visual neurons of birds, although a small number of monopolar neurons resembling those of the avian isthmo‐optic nucleus may also be observed. A few retinopetal neurons in the griseum centralis tectalis were tyrosine hydroxylase (TH) immunoreactive. Moreover, in the nuclei subcoeruleus dorsalis and isthmi parvocellularis, both ipsilaterally and contralaterally, approximately one retinopetal neuron in three (35%) was immunoreactive to nitric oxide synthase (NOS), and a slightly higher proportion (38%) of retinopetal neurons were immunoreactive for choline acetyltransferase (ChAT). Some of them contained colocalized ChAT and NOS/reduced nicotinamide adenine dinucleotide phosphate–diaphorase. Fibers immunoreactive to TH, serotonin (5‐HT), neuropeptide Y (NPY), or Phe‐Met‐Arg‐Phe‐amide (FMRF‐amide) were frequently observed to make intimate contact with rhodamine‐labeled retinopetal neurons. These findings are discussed in relation to previous results obtained in other reptilian species and in birds. J. Comp. Neurol. 468:65–85, 2004.

An immunohistochemical study of putative neuromodulators and transmitters in the centrifugal visual system of the quail (Coturnix japonica)

The aim of the present study was to analyze the neurochemical properties of the centrifugal visual system (CVS) of the quail using an immunohistochemical approach by testing 16 neuropeptides (angiotensin: ANG, bradykinin: BK, cholecystokinin, dynorphin, L and M-enkephalin, beta-endorphin: beta-END, galanin, alpha-neoendorphin, neurokinin A, neuropeptide Y (NPY), ocytocin, somatostatin, substance P, vasopressin, vasoactive intestinal polypeptide) and three neurotransmitters or their synthetic enzymes (choline acetyltransferase: ChAT, tyrosine hydroxylase: TH, serotonin: 5-HT and nitric oxide synthase: NOS, including the histochemical nicotinamide adenine dinucleotide phosphate diaphorase technique). For each substance, the somatic and afferent fiber and terminal labeling was analyzed within the nucleus isthmo-opticus (NIO) and the ectopic area (EA) and compared with that of retinopetal cell bodies labeled retrogradely with RITC following its intraocular injection (double-labeling procedure). The results showed that none of the centrifugal neurons were reactive to any of the substances tested. In contrast, all with the exception of ANG, BK and beta-END, labeled fibers and terminals within the EA and only four (ChAT, 5-HT, NPY and NOS) within the NIO. Possible sources of these immunoreactive fibers terminating in the NIO and EA were investigated by mapping the somatic immunolabeling of the different substances within brainstem regions previously shown by Miceli and other authors to project upon the centrifugal neurons. The data suggests that, besides the rapid retino-tecto-NIO-retinal loop, which facilitates the transfer of meaningful or more relevant information within particular portions of the visual field, the multiple afferent input which stems from various brainstem regions utilizes a wide range of neuroactive substances. Some of these afferent projections upon the centrifugal neurons appear to belong to nonspecific systems which might play a role in modulating the excitability of centrifugal neurons as a function of arousal.

Serotonin immunoreactivity in the retinal projecting isthmo-optic nucleus and evidence of brainstem raphe connections in the pigeon.

Serotonin (5-HT) immunoreactive (-ir) profiles within the isthmo-optic nucleus (ION) of the centrifugal visual system (CVS) were studied in the pigeon using light microscopic immunohistofluorescent and electron microscopic immunocytochemical pre-embedding techniques. The brainstem origin of the 5-HT input upon the ION was determined by combining 5-HT immunohistofluorescence (FITC) and retrograde transneuronal tracing after intraocular injection of Rhodamine beta-isothiocyanate. The light microscopic results showed that 5-HT endings were mainly localised within the neuropillar zones of the ventral ION. The 5-HT-ir cell bodies, belonging to a lateral extension of the dorsal raphe system, were observed within the same region as the centrifugal ectopic neurons (EN) underlying the ION and some displayed dendritic processes which penetrated the nucleus. Double-labeled neurons, representing 5-HT-ir afferents to the ION, were identified only within the n. linearis caudalis region of the ventral raphe. The electron microscopic results confirmed the presence of 5-HT-ir dendritic processes within the ventral part of the nucleus and showed that they were contacted by axon terminals belonging to intrinsic interneurons. The functional organisation of the ION and the possible contribution of serotonergic raphe afferents and efferents are discussed in relation to present hypotheses linking the avian CVS to mechanisms of visual attention.

The primary visual system of flatfish: an evolutionary perspective.

SummaryThe retinal projections of two species of flatfish (Scophthalmus maximus, Scophthalmidae; Platichthys flesus, Pleuronectidae) were investigated by autoradiography and by a HRP technique. Contralateral projections to five hypothalamic centres (area optica preoptica ventralis, nucleus opticus preopticus parvocellularis posterior pars lateralis, n. suprachiasmaticus, n. opticus hypothalami ventromedialis and area optica hypothalami posterior), thirteen thalamo-pretectal centres (nucleus opticus dorsolateralis (partes medialis, ventralis and lateralis), n. opticus ventrolateralis, n. opticus commissurae posterioris (partes dorsalis and ventralis), n. opticus accessorius, n. geniculatus lateralis mesencephali, nn. opticus pretectalis dorsalis, medialis and ventralis and n. corticalis), three layers of the optic tectum (stratum opticum pars externa, stratum fibrosum et griseum superficiale, stratum album centrale), and a single target in the tegmentum (n. opticus tegmenti mesencephali dorsalis), were identified in both species. Interspecific variation of the contralateral visual projections is relatively small. Ipsilateral visual projections of fibres which recross the midline in the minor and transverse commissures were also identified; in S. maximus this ipsilateral contingent is poorly developed and concerns principally hypothalamic structures, while in P. flesus the ipsilateral projections are considerably more extensive and involve both hypothalamic and thalamo-pretectal primary visual centres. No differences in the projections from the fixed and from the migrated eye were observed in either species. The findings are discussed in the general context of the existing literature on the visual projections of teleosts, in an attempt to characterize the primary visual system of the Pleuronectiformes in an evolutionary context.